DNA replication of the parvovirus Kilham rat virus. I. Characterization of intracellular forms of viral DNA extracted by guanidine hydrochloride
The following experimental procedures are described: infection of rat kidney cell cultures with Kilham rat virus (KRV); labeling and extraction of DNA; sucrose density gradient sedimentation; digestion by S/sub 1/ nuclease; and chromatography of DNA on benzoylated-napthoylated DEAE(BND)-cellulose columns. Results are discussed with regard to isolation and purification of intracellular forms of KRV DNA; velocity sedimentation in alkaline sucrose gradients; S/sub 1/ nuclease digestion of intracellular viral DNA; BND-cellulose chromatography; and pulse-chase experiment for identification of RIs. Results indicate that within the infected cell and at a time of active viral DNA replication, three populations of viral-specific molecules can be obtained by extraction with guanidine HCl. Characteristics of the three populations are described. (HLW)
- Research Organization:
- Tennessee Univ., Oak Ridge (USA). School of Biomedical Sciences; Oak Ridge National Lab., TN (USA)
- DOE Contract Number:
- W-7405-ENG-26
- OSTI ID:
- 7100340
- Report Number(s):
- CONF-770555-2
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
59 BASIC BIOLOGICAL SCIENCES
ANIMAL CELLS
ANIMALS
BIOCHEMISTRY
BODY
CARBONIC ACID DERIVATIVES
CHEMISTRY
CHROMATOGRAPHY
DNA
ENZYMES
EXTRACTION
GUANIDINES
INFECTIVITY
KIDNEYS
MAMMALS
MICROORGANISMS
NUCLEASES
NUCLEIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
ORGANS
PARASITES
PHOSPHOTRANSFERASES
RATS
RODENTS
SEDIMENTATION
SEPARATION PROCESSES
TRANSFERASES
VERTEBRATES
VIRUSES